Under this Phase I SBIR, Lumiphore will use its proprietary lanthanide technology to develop the `next-generation'of multiplex capable reporter dyes with photophysical properties superior to those currently available. Lumiphore's lanthanide compounds have longer life-times, a larger Stokes shift, and sharper fluorescent peak profiles that conventional organic fluorescent materials, and have greater stability and brightness than commercially available lanthanide- based fluorescence products. Under this SBIR effort, Lumiphore proposes to develop Lumi4"- Tb (Donor) for multi-color time resolved fluorescence. Lumi4"-Tb, originally developed at University of California - Berkeley, is an isophthalamide based macrocyclic chelate that has an exceptionally high quantum yield (60%), extinction coefficient (26,000 M-1cm-1), and long lifetime (2.7 ms) at dilute concentrations in standard aqueous buffer environments. Lumi4TM-Tb has four narrow emission peaks centered at approximately 490nm, 545nm, 590nm, and 620nm. These emission peaks form the basis of the proposed multicolor fluorescent system through energy transfer coupling to a wide range of conventional organic fluorophores (Acceptors) with overlapping excitation maxima. We propose to also vary the linker characteristics between Donor and Acceptors such that emission lifetimes of otherwise identical pairs are discernable. This will create an additional multiplex property in addition to emission wavelength. The proposed research will overcome fundamental limitations of organic fluorescent dyes that have limited the sensitivity, speed, and multiplexing capability and reliability of florescence applications for high throughput screening (HTS) drug discovery, genomic screening, rapid DNA sequencing, clinical diagnostics and fluorescence microscopy. PUBLIC HEALTH RELEVANCE: The proposed research will impact public health by enabling highly multiplexed diagnostic assay formats that combine fast turnaround times and high assay throughput efficiency. These assays will be used in high throughput screening (HTS) drug discovery, genomic screening, rapid DNA sequencing, clinical diagnostics and fluorescence microscopy applications for both basic and applied biomedical research.